دورية أكاديمية
抗血小板膜糖蛋白Ibα胞内段多肽多克隆抗体的制备及初步应用 ; Preparation and preliminary application of rabbit polydonal antibodies against intracellular peptides of human platelet glycoprotein GPIbα
| العنوان: | 抗血小板膜糖蛋白Ibα胞内段多肽多克隆抗体的制备及初步应用 ; Preparation and preliminary application of rabbit polydonal antibodies against intracellular peptides of human platelet glycoprotein GPIbα |
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| المؤلفون: | 元艳宏, 康佳蕊, 闫荣, 程虹, 庄逢源, 王月丹, 戴克胜 |
| المساهمون: | 100083,北京航空航天大学生物与医学工程学院血液与细胞生物学实验室, 中国航天员科研训练中心分子和细胞研究室, 北京大学医学部免疫系, 北京航空航天大学生物与医学工程学院血液与细胞生物学实验室,100083 |
| المصدر: | 万方 ; http://d.g.wanfangdata.com.cn/Periodical_zhyx200912010.aspxTest |
| بيانات النشر: | 中华医学杂志 |
| سنة النشر: | 2009 |
| المجموعة: | Peking University Institutional Repository (PKU IR) / 北京大学机构知识库 |
| مصطلحات موضوعية: | 血小板, 血小板糖蛋白GPIb-IX复合物, 磷酸丝氨酸, 抗体, Blood platelets, Platelet glycoprotein GPIb-IX complex, Phosphoserine, Antibodies |
| الوصف: | 目的 制备兔抗人血小板膜糖蛋白(GP)Ibα C端557～561氨基酸序列多肽的多克隆抗体,并初步用于人血小板GPIbα C端559位丝氨酸磷酸化状态的检测.方法 应用化学方法合成C-R-G-S-L-P(559位丝氨酸非磷酸化,Ser559)和C-R-G-s(p)-L-P(559位丝氨酸磷酸化,pSer559)多肽.将2种多肽分别与钥孔蜮血蓝蛋白交联后,以皮下注射法分别免疫新西兰大白兔,分离获得2种抗血清(抗Ser559多抗和抗pSer559多抗).应用斑点印迹和酶联免疫吸附试验(ELISA)方法 对抗血清进行鉴定并检测效价.从人血小板裂解液中分离纯化血小板GPIbα,利用抗Ser559多抗和抗pSer559多抗、采用ELISA方法检测人血小板GPIbαC端559位丝氨酸磷酸化状况.结果 所制备的2种多抗分别特异性识别各自抗原,效价分别为1:32 000和1:64 000.2种多抗均可与纯化的人血小板GPIbα特异结合,表明人血小板GPIbα 559位点丝氨酸存在磷酸化与非磷酸化两种状态.结论 应用人工合成多肽成功制备出2种可特异性识别丝氨酸磷酸化状态的兔抗人血小板GPIbα胞内段多肽多抗,并证明人血小板GPIbαC端559位丝氨酸存在磷酸化状态. ; Objective To prepare rabbit polyclonal antibodies against intraceUular peptides of human platelet glycoprotein GPIbα. Methods Two peptides corresponding to human platelet GPIbα C-terminus were synthesized and purified by high-performance liquid chromatography (HPLC). The peptides were cross-linked with keyhole limpet hemocyanin (KLH). Two New Zealand white rabbits were immunized with conjugated peptides for 3 times. The polyclonal antibodies were purified by Ammonium Sulfate Precipitation and identified by dot blotting and ELISA. GPIbot intracellular poptides phospharylation was tested with these polyclonal antibodies by ELISA. Results The titers of the two polyclonal antibodies against the GPIbα C-terminus poptides were 1 : 32 000 and 1 : 64 000 respectively and both of these antibodies reacted with purified GPlbot. Conclusions Two rabbit polyclonal antibodies against C-terminal peptides of human platelet GPIbα have been prepared successfully, providing a way for the preparation of these kinds of antibody. Both phosphorylation and dephosphorylation states exist in the intracellular poptide of human platelets. ; 国家自然科学基金; 教育部新世纪优秀人才资助计划; 全国优秀博士论文获得者专项基金 ; 中文核心期刊要目总览(PKU) ; 中国科技核心期刊(ISTIC) ; 中国科学引文数据库(CSCD) ; 0 ; 12 ; 826-830 ; 89 |
| نوع الوثيقة: | journal/newspaper |
| اللغة: | Chinese |
| تدمد: | 0376-2491 |
| العلاقة: | 中华医学杂志.2009,89,(12),826-830.; 1277496; http://hdl.handle.net/20.500.11897/290003Test |
| DOI: | 10.3760/cma.j.issn.0376-2491.2009.12.010 |
| الإتاحة: | https://doi.org/20.500.11897/290003Test https://doi.org/10.3760/cma.j.issn.0376-2491.2009.12.010Test https://hdl.handle.net/20.500.11897/290003Test |
| رقم الانضمام: | edsbas.AE31903D |
| قاعدة البيانات: | BASE |
Full Text Finder | تدمد: | 03762491 |
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| DOI: | 10.3760/cma.j.issn.0376-2491.2009.12.010 |